A neuron-specific gene transfer by a recombinant defective Sindbis virus

We examined the possibility that Sindbis virus, an alpha virus with a singl e-stranded RNA genome, would be applied for neuronal gene transfer. The rec ombinant defective Sindbis viruses were constructed by replacing the struct ural genes of Sindbis virus with genes encoding beta-galactosidase (rdSind- lacZ) or enhanced green fluorescent protein (rdSind-EGFP). In neuron-glia c ocultures prepared from the neocortex, hippocampus, and striatum, EGFP or b eta-galactosidase was expressed selectively in neurons 24 h after infection with rdSind-EGFP or rdSind-lacZ. Most cortical neurons were infected with rdSind-lacZ at a multiplicity of infection (M.O.I.) of 5 while glial cells were little infected. In addition, transient neuron-specific expression of beta-galactosidase was observed near injection sites over the next 3 d foll owing administration of rdSind-lacZ in adult rat. In the cortical neurons i nfected with rdSind-EGFP, treatment with NMDA induced neuritic blebs and ce ll body swelling in a Na+-dependent manner. Therefore, recombinant defectiv e Sindbis viruses can be used as an efficient and selective vector for gene transfer into neurons and applied to investigate biological role of target genes delivered into neurons in vitro and in vivo. (C) 1998 Elsevier Scien ce B.V. All rights reserved.